Novel flow cytometry-based assay for HIV-infected cells

基于流式细胞术的新型 HIV 感染细胞检测方法

• 批准号: 8656014
• 负责人: JOSEPH M MCCUNE
• 金额: $ 22.06万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-12-01 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8656014
• 关键词: Acquired Immunodeficiency Syndrome; Age; Biological Assay; Biological Models; Blood; CD4 Positive T Lymphocytes; Cardiovascular Diseases; Cell Count; Cell Line; Cells; Cellular Assay; DNA; Detection; Development; Disease; Disease Progression; Effectiveness; Epidemic; Evaluation; Event; Flow Cytometry; Fluorescent in Situ Hybridization; Frequencies; General Population; HIV; HIV Infections; History of Medicine; In Vitro; Individual; Infection; Intervention Trial; Intestines; Investigation; Label; Location; Measurement; Measures; Memory; Methods; Modeling; Monitor; Neurocognitive; Nucleic Acids; Patients; Pattern; Phase; Phenotype; Population; Process; Production; RNA; Recovery; Resolution; Sampling; Sensitivity and Specificity; Signal Transduction; Specificity; Specimen; Subgroup; Testing; Time; Tissues; Viral; Virus; Virus Latency; Virus Replication; antiretroviral therapy; assay development; base; clinically relevant; cost; design; high throughput screening; in vitro Model; in vivo; killings; macrophage; novel; novel therapeutics; population based; prevent; public health relevance; reactivation from latency; research study; screening

DESCRIPTION (provided by applicant): Antiretroviral therapy (ART) for HIV infection is one of the most important and rapid advances in the history of medicine. However, while effective at limiting virus replication and slowing the course of disease, ART cannot be stopped without rapid rebound of virus and progression of disease. In addition, while AIDS is prevented in patients on ART, significant non-AIDS events such as cardiovascular disease and neurocognitive decline occur more frequently and at an earlier age in HIV-infected subjects on ART than in the general population. Finally, the epidemic is too large and the barriers to roll-out too high to effectively treat every infected individual to end the epidemic. It is therefore imperative that we understand the mechanisms of how HIV persists in the infected individual to design novel therapies to eradicate the virus or functionally cure the patient. The first step in tis understanding is the development of assays to measure the level of virus persistence. Several assays exist, but have serious limitations in terms of how much information is provided, how quickly they can be performed, how expensive they are, and how variable the measurement is. We propose here a novel method to detect HIV-infected cells at single cell resolution using flow cytometry. This method, once optimized, will provide more information, be performed more rapidly, cost less, and have less variability than other assays for the measurement of HIV reservoirs. We will optimize this assay and then apply it to understand the state of the HIV reservoir in various groups of HIV-infected subjects to evaluate whether some clinically different subjects may have different sizes, activities, or reactivation potentials of their HIV reservoirs ad also to develop a high-throughput screening assay for the evaluation of novel therapeutics to reactivate or kill the latent reservoir. This process will demonstrate the utility of this assay an highlight situations in which it can be applied as well as disseminate the assay for use by others to apply as they see fit.

描述（由申请人提供）：针对 HIV 感染的抗逆转录病毒疗法 (ART) 是医学史上最重要和最快速的进步之一。然而，虽然 ART 可以有效限制病毒复制和减缓病程，但如果病毒不迅速反弹和疾病进展，就无法停止 ART。此外，虽然接受抗逆转录病毒治疗的患者可以预防艾滋病，但与普通人群相比，接受抗逆转录病毒治疗的艾滋病毒感染者发生心血管疾病和神经认知能力下降等重大非艾滋病事件的频率更高，发生的年龄也更早。最后，疫情太大，推广存在障碍 太高了，无法有效治疗每个感染者以结束流行病。因此，我们必须了解艾滋病毒在感染者体内持续存在的机制，以设计新的疗法来根除病毒或功能性治愈患者。理解这一点的第一步是开发测量病毒持久性水平的测定方法。存在多种测定方法，但在提供的信息量、执行速度、成本以及测量的可变性方面存在严重限制。我们在这里提出了一种使用流式细胞术以单细胞分辨率检测 HIV 感染细胞的新方法。该方法一旦优化，将提供更多信息，执行速度更快，成本更低，并且与其他测量 HIV 病毒库的方法相比，变异性更小。我们将优化该测定，然后应用它来了解不同组的 HIV 感染受试者中 HIV 储存库的状态，以评估一些临床上不同的受试者是否可能具有不同的 HIV 储存库大小、活动或重新激活潜力，并开发一种高通量筛选测定法，用于评估重新激活或杀死潜在储存库的新疗法。此过程将展示该测定法的实用性，突出显示其可以应用的情况，并传播该测定法以供其他人在他们认为合适的情况下使用。